Testing Plant Substances as Potential Medicines

Background:
We went to BioMarin and learned about different medicines and drugs they are creating. This
lab is meant to expand on what we learned there and what we learned in class about medicine
from plants.
Purpose/ Objective:
The purpose of the lab is to determine if the plant material that we brought in is able to be
used as a medicine. We find this by seeing if the plants contain active ingredients that inhibit the
growth of bacteria.
Materials:
Pointsettia (plant)

E. Coli JM109 (stock plate)

10 mL syringe

10 mL pipet

test tubes

100 mL beakers

Methanol

5mm filter paper

1 mL pipet

250 mL media bottle

Ampicillin

LB Agar

Inoculating Loop

LB broth base

60x15 Petri dish

Plastic funnels

Procedure:
1. Measure 2 grams of plant material.
2. Using a mortar and plastic, grind up 2 grams of plant tissue with 10 mL of
deionized water.
3. Let sit for 3 minutes.
4. Filter the sample through an 11 cm filter paper funnel.
5. Filter sterilized the extract using a syringe filter.
6. Collect 1 mL of extract into a 1.7 mL microtube. Label the tube.
7. Repeat the steps 1-6 using methanol instead of water.
Filter Sterilization Procedure:
1. Attach pre-fitter to syringe and rinse with H2O
2. Carefully open sterilization filter, keeping the filter in its plastic covering
3. Load approx. 1.5 ml of H2O-based filtrate using a pipet
4. Depress the plunger, collecting the sterile-filtered filtrate into microfuge tube
5. Quickly close microfuge cap until a snap sound is heard
6. Use Flame -Sterilized Forceps to transfer filter squares (2 per group) into H2O
control and ampicalli and control microtubes
7. Label plate into four grids
8. Ms. Flasher will dispense 1 mL E-coli in each plate
9. Flame-Sterilize Spreading Loop and spread bacteria around

10. Place filter disks in plate

Results:
24 hours

48 hours

72 hours

plate 1

no signs of
contamination

slight antimicrobial
ring

more noticeable
antimicrobial ring

plate 2

signs of
contamination

slight antimicrobial
ring

more noticeable
antimicrobial ring

Day 1:

Day 2:

Day 3:

Analysis:
With our water petri dish there was a clearing around the ampicillin. Ampicillin is an
antimicrobial compound that causes death of bacterium E. coli by inhibiting the cell wall
synthesis. The bacterial lawn didn't grow right up to the negative control disk. (That's where the
clearance was located.) Water would not be expected to have antimicrobial activity, which
makes it a good negative control for this experiment. I didn't see any clearings around the disks
soaked in plant extract. There was only a clearing around the ampicillin. A possible problem that
might have affected the results is that something wasn't sterile. I thought that this lab would
have been much more effective if we did it over a shorter period of time to eliminate
contamination possibilities.